• 제목/요약/키워드: Dendrolimus punctatus tetravirus

검색결과 1건 처리시간 0.014초

Expression and Characterization of RNA-dependent RNA Polymerase of Dendrolimus punctatus Tetravirus

  • Zhou, Liang;Zhang, Jiamin;Wang, Xiaochun;Jiang, Hong;Yi, Fuming;Hu, Yuanyang
    • BMB Reports
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    • 제39권5호
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    • pp.571-577
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    • 2006
  • Dendrolimus punctatus tetravirus (DpTV) has been identified as a new member of the genus Omegatetravirus of the family Tetraviridae that may be related serologically to Nudaurelia capensis virus ($N{\omega}V$). To establish the function of DpTV RNA genome and to better understand the mechanism of viral replication, the putative RNA-dependent RNA polymerase (RdRp) domain has been cloned and expressed in Escherichia coli. The recombinant protein was purified on a Ni-chelating HisTrap affinity column and demonstrated to initiate viral RNA synthesis in a primer-independent manner but not by terminal nucleotidyle transferase activity in the presence of $Mg^{2+}$ and RNA template. Mutation of the GDD to GAA interferes with the residues at the polymerase active site and metal ions, and thus renders the polymerase inactive.